Use of lectin binding characteristics to identify gastrointestinal parasite eggs in faeces

Abstract

Identification of the presence and abundance of species is important when choosing therapies and control strategies for internal parasitism of livestock. Here we examine lectin binding characteristics of eggs isolated from sheep faeces as a means for identifying the parasite genera contributing to infection. The intensity of lectin staining varied with incubation time, incubation volume, concentration of lectin and concentration of eggs. Formalin fixed eggs had greater autofluorescence but exhibited the same lectin staining pattern as fresh eggs. The stage of egg development did not influence staining. Eggs from Haemonchus contortus, H. placei, Trichostrongylus colubriformis, T. vitrinus, Ostertagia circumcincta, Nematodirus spathiger and the cestode Monezia expansa were incubated with a panel of fluoroscein isothiocyanate (FITC)-labelled lectins. Lectin binding exhibited a genus specific pattern. Haemonchus spp. stained strongly positive with peanut agglutinin (PNA), and were positive for concanavalin A (ConA), Ricinus communis agglutinin (RCA) and Maclura pomifera lectin (MPA). Trichostrongylus spp. were PNA−, ConA−, RCA− and strongly MPA+. O. circumcincta were weakly positive for PNA, MPA, ConA and negative for RCA. N. spathiger were weakly positive for the four lectins, and M. expansa were weakly positive for PNA, RCA and MPA and were strongly ConA+. The genus specificity of lectin staining was used to identify the presence of Trichostrongylus and Haemonchus eggs in faeces from sheep with mixed field infections, however correspondence between lectin staining and larval differentiation for identifying a low prevalence of Ostertagia in the field infection was poor. Refinements in methods for rapid egg isolation may improve egg differentiation on the basis of lectin staining, which could be undertaken by flow cytometry or microscopy.